Advances in analysis of novel protein therapeutics and clinical samples with LC-MS/MS methods

Abstract

Analytical characterization is essential for drug efficacy, stability and safety. Significant concern in protein stability and potential immunogenicity requires detailed characterization of protein therapeutics. Therefore, the ability to detect product variants from non-clonal cell lines is very important. To fulfill this job, LC-MS/MS becomes the perfect choice. This Ph.D. research discusses advances in analysis of novel protein therapeutics and clinical samples with LC-MS/MS methods. Recombinant protein therapeutics is a major focus of my research work. In Chapter 1, an overview of production of recombinant protein therapeutics and the potential heterogeneity, in particular post-translational modifications, is reviewed. Common techniques for protein characterization are discussed, including electrophoresis, chromatography, and mass spectrometry. Because the major focus of my research is about mass spectrometry, further details about ionization method, mass analyzer, hybrid mass spectrometer, fragmentation method and LC-MS/MS analysis are reviewed. In addition, common strategies for protein characterization are discussed. Furthermore, biology part for proteomic analysis is reviewed. ERBB2, an important oncogene, is of particular interest and its association to cancer disease is included.